مطالعه کارآیی روش تشخیصی آزمایش مستقیم میکروبی، PCR900IS و کشت میکروبی در تشخیص باکتری مایکوباکتریوم اویوم تحت گونه پاراتوبرکلوزیس در مدفوع گاوهای به ظاهر سالم

محورهای موضوعی : آسیب شناسی درمانگاهی دامپزشکی

یونس انزابی ¹ (گروه پاتوبیولوژی، دانشکده دامپزشکی، دانشگاه آزاد اسلامی واحد تبریز، تبریز، ایران)
صمد فراشی بناب ² (گروه میکروبیولوژی و ایمونولوژی، دانشکده دامپزشکی، دانشگاه تهران، تهران، ایران)
غلامعلی مقدم ³ (گروه علوم درمانگاهی، دانشکده دامپزشکی، دانشگاه تبریز، تبریز، ایران)

تاریخ دریافت : 1387/10/29 تاریخ پذیرش : 1388/01/31 تاریخ انتشار : 1387/12/01

کلید واژه: PCR, یماری یون, آزمایش مستقیم میکروبی, کشت مدفوع, مایکوباکتریوم اویوم تحت‌گونه پاراتوبرکلوزیس,

چکیده مقاله :

بیماری یون یا پاراتوبرکلوزیس نوعی آنتریت گرانولوماتوزی مزمن در نشخوارکنندگان با عامل مسبب مایکوباکتریوم اویوم تحت گونه پاراتوبرکلوزیس (MAP) می باشد و خسارات اقتصادی فراوانی به صنعت دامپروری به‌خصوص گاوداری شیری در سراسر دنیا وارد می کند. تشخیص MAP در نمونه های بالینی با روش کشت میکروبی به عنوان روش استاندارد طلایی به 16-6 هفته زمان نیاز دارد. از طرف دیگر شناسایی سریع و دقیق گاوهای دفع کننده میکروب فوق در مدفوع برای کنترل موفق بیماری در گله لازم است. در این تحقیق، بر روی مدفوع 100 رأس گاو به ظاهر سالم از گاوداری های صنعتی تبریز با سابقه بیماری یون، آزمایش مستقیم میکروبی با رنگ آمیزی ذیل نلسن، کشت میکروبی در محیط زرده تخم مرغ هرولد و دو روش direct PCRبر پایه عنصر 900IS انجام شد. در آزمایش مستقیم میکروسکوپی 7 نمونه (7 درصد)، در کشت میکروبی 14 نمونه (14 درصد)، در PCR با جفت پرایمر 91F/90F 15 نمونه (15 درصد) و در PCR با جفت پرایمر 26FP/25FP 25 نمونه (25 درصد) مثبت ثبت شدند. این نتایج نشان داد روش PCR تعداد موارد مثبت بیشتری را شناسایی می کند، بنابراین می توان از آن برای شناسایی سریع و در عین حال دقیق تر گاوهای دفع کننده MAP در مدفوع استفاده کرد. هم‌چنین نوع پرایمر در حساسیت تست PCR نقش مهمی ایفا می کند.

چکیده انگلیسی:

     Johne’s disease or paratuberculosis is a chronic granulomatous enteritis in ruminants caused by Mycobacterium avium subsp. paratuberculosis (MAP). The disease is responsible for significant economic losses in dairy industry worldwide.  Microbial culture as a golden standard test for detection of MAP in faecal specimens requires 6-16 weeks to complete, whereas accurate and rapid identification of cattle shedding MAP in their feces is essential for successful control of the disease in herds. In the present study, direct microbial diagnosis by Ziehl-Neelsen acid-fast staining, microbial culture on Herrolds’ egg yolk media and two IS900 direct PCR assays were carried out on 100 fecal specimens of apparently healthy cattle collected from dairy herds of Tabriz with a history of Johne’s disease. The number of positive specimens identified by the direct microbial diagnosis, microbial culture and PCR with F90/F91 and FP25/FP26 primes were 7 (7%), 14 (14%), 15(15%) and 25(25%) respectively. These results indicated that PCR detected more positive cases therefore it can be employed for rapid and accurate diagnosis of cattle shedding MAP it their feces and the type of primer used has a significant role in the sensitivity of this test. Direct microbial diagnosis by Ziehl-Neelsen acid-fast staining identified 7 (7%) specimens, two IS900 direct PCR assays identified 15 (15%) and 25 (25%) specimens, respectively, and microbial culture identified 7 (7%) specimens as positive. Collectively, these data indicate that PCR detection of MAP was more sensitive than direct microbial diagnosis by Ziehl-Neelsen acid-fast staining or faecal culture, especially if appropriate primers were used.

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